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Purpose To assess the noninferiority of a single platelet-rich plasma (PRP) injection compared with hyaluronic acid (HA), to alleviate pain and enhance functional capacity in knee osteoarthritis, and identify biological characteristics of PRP that may affect their efficacy. Methods Fifty-four patients with symptomatic knee osteoarthritis received a single injection of either PRP (26 patients) or HA (28 patients). They were assessed at baseline and at 1, 3, and 6 months. The primary endpoint was the change in Western Ontario and McMaster Universities Arthritis Index (WOMAC) score at 3 months, and secondary endpoints were responders' rate (improvement of at least 5 points or 40% of WOMAC total score at 3 months) of pain evaluation and patient's subjective satisfaction. Cell counts and the contents of vascular endothelial growth factor (VEGF), platelet-derived growth factor–AB (PDGF-AB), transforming growth factor beta 1 (TGF-β1) content of injected PRP were assessed to analyze their relationship with clinical outcome. Results Both treatments proved their improvement in knee functional status and symptom relief, with a significant decrease observed at 1 month on all scores except for pain VAS in PRP group and WOMAC function score in the HA group. No difference between groups regarding WOMAC and VAS scores was observed. A higher percentage of responders was observed in the PRP group (72.7%) than in the HA group (45.8%) without significance ( P = .064). The quantity of injected PDGF-AB and TGF-β1 correlated with the change in WOMAC scores at 3 months and was lower in responders than in nonresponders ( P = .009 and P = .003, respectively). Conclusions Current results indicated that a single injection of very pure PRP offers a significant clinical improvement in the management of knee osteoarthritis, equivalent to a single HA injection in this patient population. Moreover, a significant correlation between the doses of TGF-β1 and PDGF-AB and the worsening of WOMAC score 3 months after the procedure was found. Level of Evidence Level II, randomized double blind controlled trial.

International audience; Platelet-Rich Plasma (PRP) is an autologous biological therapy obtained by centrifuging the patient's own blood to concentrate platelets. The addition of autologous thrombin and calcium chloride to PRP allows the production of a semi-solid form called PRP gel. PRP gel is increasingly used in a variety of tissue defects and predominantly in the management of non-healing chronic wounds. The topical application of PRP gel seems promising due to the capability of platelets to store and secrete growth factors (GF), fibrin and cytokines, which are essentials for wound healing. Most patients who suffered from chronic wounds are elderly patients with comorbidities and polypharmacy including antithrombotic drugs such as antiplatelet agents (AP) or anticoagulants (AC), which could hamper the feasibility of this autologous platelet-derived therapy. To date, no study has investigated PRP gel formation in patients with AP or AC. The aim of this study was to evaluate the influence of AP or AC drugs on the production of PRP gel formation from elderly patients. Different biological characteristics were determined to qualify the production of PRP gel from such patients (Interquartile range (IQR) = 75-92 years) compared to healthy volunteers (IQR = 23-37 years). No significant difference was observed in the volume, composition (quantity of platelets, leukocytes and red blood cells) and functionality of platelets from PRP except a higher ADP-induced P-selectin expression in healthy donors compared with elderly patients. Autologous thrombin characteristics were similar in the two groups. Gel time formation (IQR: 120-195 seconds for controls and 135-210 seconds for elderly patients) and final composition of PRP gel were not significantly modified. Concentrations of theoretical thrombin generated in the serum and in the gel were inversely correlated with the time of formation of PRP gel (r(2) = 0.57, p = 0.012). Altogether these data indicate that PRP gel preparation is not impacted by the use of antithrombotic drugs. Such results support the feasibility of using this innovative autologous biotherapy in the management of elderly patients with non-healing chronic wounds.

The disruption of endothelial homeostasis is a major determinant in the pathogenesis of systemic sclerosis (SSc) and is reflected by soluble and cellular markers of activation, injury and repair. We aimed to provide a combined assessment of endothelial markers to delineate specific profiles associated with SSc disease and its severity. We conducted an observational, single-centre study comprising 45 patients with SSc and 41 healthy control subjects. Flow cytometry was used to quantify circulating endothelial microparticles (EMPs) and CD34+ progenitor cell subsets. Colony-forming unit-endothelial cells (CFU-ECs) were counted by culture assay. Circulating endothelial cells were enumerated using anti-CD146-based immunomagnetic separation. Blood levels of endothelin-1, vascular endothelial growth factor (VEGF) and soluble fractalkine (s-Fractalkine) were evaluated by enzyme-linked immunosorbent assay. Disease-associated markers were identified using univariate, correlation and multivariate analyses. Enhanced numbers of EMPs, CFU-ECs and non-haematopoietic CD34+CD45− endothelial progenitor cells (EPCs) were observed in patients with SSc. Patients with SSc also displayed higher serum levels of VEGF, endothelin-1 and s-Fractalkine. s-Fractalkine levels positively correlated with CD34+CD45− EPC numbers. EMPs, s-Fractalkine and endothelin-1 were independent factors associated with SSc. Patients with high CD34+CD45− EPC numbers had lower forced vital capacity values. Elevated s-Fractalkine levels were associated with disease severity, a higher frequency of pulmonary fibrosis and altered carbon monoxide diffusion. This study identifies the mobilisation of CD34+CD45− EPCs and high levels of s-Fractalkine as specific features of SSc-associated vascular activation and disease severity. This signature may provide novel insights linking endothelial inflammation and defective repair processes in the pathogenesis of SSc.

Place: United States

Le consortium TOURNESOL, regroupant 11 equipes de chercheurs et de cliniciens de trois villes (Marseille, Montpellier et Rouen) et finance par la Fondation de l’Avenir, a pour mission de mener deux essais cliniques destines a reparer des nerfs peripheriques leses–nerfs facials et des membres – grâce a des greffes autologues de cellules souches nasales. Pourquoi la therapie cellulaire ? Environ 100 000 operations de reparation nerveuse sont realisees chaque annee aux Etats-Unis et en Europe. Elles representent 2,8 % des traumatismes et sont le plus souvent responsables d’une perte de fonction voire d’invalidite. Malgre des progres dans le delai de prise en charge et dans la qualite du geste technique chirurgical, il n’y a eu aucun progres majeur depuis une vingtaine d’annees. Une approche de therapie cellulaire pourrait limiter l’inflammation et ameliorer la croissance axonale. Pourquoi les cellules souches olfactives nasales ? Chez le mammifere adulte, y compris chez l’homme, la muqueuse olfactive est le seul tissu nerveux capable d’auto-renouvellement. La plasticite de ce tissu est en partie assuree par des cellules souches qui sont multipotentes. Comme demontre par plusieurs membres du consortium, elles peuvent donner naissance a des cellules neurales (neurones, astrocytes, oligodendrocytes, cellules de Schwann). Leur potentiel therapeutique a ete teste avec succes dans differents modeles animaux de paraplegie, de la maladie de Parkinson, de surdite et d’amnesie. Que disent les etudes menees chez le rat par le consortium. Deux etudes, menees en parallele sur le nerf facial et le nerf peronier ont permis de montrer que la greffe de cellules souches olfactives : ameliore la recuperation fonctionnelle de la mobilite faciale ; diminue les syncinesies œil-bouche ; accroit les capacites locomotrices ; ameliore le seuil tetanique, le temps de contraction et le taux de relaxation musculaire Les cellules souches olfactives, medicament de therapie innovante, agree par l’ANSM. Avant de mener les essais cliniques, il a ete necessaire de cultiver les cellules souches dans un laboratoire de therapie cellulaire, dans des conditions repondant aux normes europeennes. Les essais cliniques multicentriques porteront sur une premiere serie de 10 patients avec des lesions negligees des nerfs collateraux et de la branche sensitive du nerf radial avec une perte de substance. La periode d’evaluation post-greffe s’etalera sur 12 mois. La chirurgie regenerative va representer une revolution dans la reparation des membres. Et les nerfs peripheriques devraient en beneficier.

Although the European Union merely followed the initiatives of the United States and Japan by introducing special regimes for orphan medicinal products, it has introduced a special status for a new category of biological medicinal products, advanced therapy medicinal products (ATMPs), adopting specific associated regulations. European Regulation (which constitutes the highest legal instrument in the hierarchy of European law texts) [EC] No. 1394/2007, published in 2007, uses this term to define somatic cell therapy medicinal products, tissue-engineered products, and gene therapy medicinal products, possibly combined with medical devices. The stated objective was two-fold: both to promote their industrialization and market access, while guaranteeing a high level of health protection for patients. Since publication of the regulation, few marketing authorizations have been granted in Europe, and these have not been accompanied by commercial success. However, certain recent studies show that this is a growing sector and that France remains the leading European nation in terms of clinical trials. This round table brought together a panel of representatives of French public and private protagonists from the advanced therapy sector. The discussions focused on the conditions to ensure the success of translational research and, more generally, the French advanced therapy sector. These enabled a number of obstacles to be identified, which once lifted, by means of recommendations, would facilitate the development and success of this sector.

Resume Alors que l’Union Europeenne n’avait fait que suivre les initiatives des Etats-Unis et du Japon en introduisant des regimes particuliers pour les medicaments orphelins, elle a instaure un statut particulier a une nouvelle categorie de medicaments biologiques, les medicaments de therapie innovante (MTI) et y associe des regles specifiques. Le reglement europeen (qui constitue l’instrument juridique le plus eleve dans la hierarchie des textes du droit europeen) CE n° 1394/2007, publie en 2007 definit sous ce terme les medicaments de therapie cellulaire somatique, les medicaments issus de l’ingenierie tissulaire, et les medicaments de therapie genique, eventuellement couples a des dispositifs medicaux. L’objectif affiche etait double : a la fois promouvoir leur industrialisation et leur acces au marche tout en garantissant un niveau eleve de protection sanitaire aux patients. Depuis la publication du reglement, les autorisations de mise sur le marche delivrees en Europe sont peu nombreuses et ne sont pas accompagnees par un succes commercial. Cependant, certaines etudes recentes montrent qu’il s’agit d’un secteur en croissance et que la France reste la premiere nation europeenne en termes de nombre d’essais cliniques. Cette table ronde a reuni un panel de representants d’acteurs publics et prives francais du secteur des therapies innovantes. Les discussions ont ete focalisees sur les conditions du succes de la recherche translationnelle et de maniere plus globale, de la filiere francaise des therapies innovantes. Elles ont conduit a identifier un certain nombre de verrous dont la levee, formulee sous forme de recommandations, permettrait de faciliter le developpement et le succes de cette filiere.

Purpose The purpose of this study was to compare the biological characteristics of platelet-rich plasma (PRP) obtained from 4 medical devices and a preparation developed in our laboratory using a single-donor model. Methods Ten healthy persons donated blood that was processed to produce PRP by use of 4 commercial preparation systems and a protocol developed in our laboratory. Volumes and platelet, white blood cell (WBC), and red blood cell concentrations were recorded. The platelet activation status was assessed by flow cytometry. Enzyme-linked immunosorbent assay was used to determine the concentrations of vascular endothelial growth factor, platelet-derived growth factor AB, epidermal growth factor, and transforming growth factor β1. We calculated platelet capture efficiency, relative composition, and increase factors from whole blood in platelets and WBC, as well as platelet and growth factor (GF) doses, provided from each preparation. Results Leukocyte-rich PRP was obtained with RegenPRP (RegenLab, Le Mont-sur-Lausanne, Switzerland) and the Mini GPS III System (Biomet Biology, Warsaw, IN) and provides PRP with higher proportions of red blood cells, WBCs, and neutrophils than leukocyte-poor PRP obtained with the Selphyl System (Selphyl, Bethlehem, PA), Arthrex ACP (Arthrex, Naples, FL), and the preparation developed in our laboratory. The highest platelet and GF concentrations and doses were obtained with the Mini GPS III System and the preparation developed in our laboratory. Different centrifugation protocols did not show differences in the percentages of activated platelets. Finally, a positive correlation between platelet doses and all the GFs studied was found, whereas a positive correlation between WBC doses and GFs was found only for vascular endothelial growth factor and epidermal growth factor. Conclusions In a single-donor model, significant biological variations in PRP obtained from different preparation systems were highlighted. The observed differences suggest different results for treated tissue and could explain the large variability in the clinical benefit of PRP reported in the literature. Clinical Relevance Our findings will help clinicians to choose a system that meets their specific needs for a given indication.

Autologous conditioned serum (ACS) is a recent biotherapy based on certain cytokines anti-inflammatory properties mainly used for the reduction of osteoarthritis (OA) symptoms. Here we investigated different physico-chemical factors influencing ACS purification and cytokine production. Human venous blood was incubated in the presence of different diameter beads (respectively 2.5, 3, 3.5, and 4 mm) or glass beads with different types of coating (polished or coated with CrSO4). Sera were recovered, and the concentrations of pro-inflammatory and anti-inflammatory relevant cytokines were measured using Luminex® technology. Fresh whole blood incubated for 24 h highly increased production of interleukin (IL)-6 and IL-8 cytokines. At the same time, the concentrations of IL-1β, IL-1 receptor agonist (IL-1Ra), IL-10, and tumor necrosis factor (TNF)-α were slightly induced. The highest cytokine concentrations were obtained with the exposure of whole blood to 3-mm glass beads and 3.5-mm polished beads. The minimum IL-1β/IL-1Ra ratio obtained was 3.2±1.3 after 24-h incubation without any beads. ACS has been shown to alleviate clinical symptoms of OA in clinical studies. This descriptive study demonstrated that different pro- and anti-inflammatory cytokines are present in ACS since no selective anti-inflammatory cytokines were produced based on the different protocols. Furthermore, we showed that CrSO4-treated glass beads are not necessary and that the absence of beads combined with a 24-h incubation could also lead to an enriched serum.

We would like to thank Dr Manetti for his relevant comments1 on our recent article in the Annals of Rheumatic Disease entitled ‘Molecular profile and proangiogenic activity of the adipose-derived stromal vascular fraction used as an autologous innovative medicinal product in patients with systemic sclerosis’2. We are pleased that this study has raised such a high interest in the scientific community and allows us to provide additional comments to those recently published3 4 in order to discuss the potential profibrotic profile of adipose-derived stem cells (ADSC) derived from patients with diffuse cutaneous systemic sclerosis (SSc). We agree that SSc is a rare but potentially lethal autoimmune disease that challenges the urgent need to develop novel therapeutic approaches. In this context, autologous fat grafting has been successfully used to limit SSc-associated clinical complications such as SSc-related perioral thickening and mouth opening limitation5 facial handicap6 …

Background New strategies are emerging in cord blood banking where focusing on birth clinics caring for a high number of mothers belonging to ethnic minorities could offer new possibilities for allotransplantation both for patients of European origin and for patients from ethnic minorities or mixed ancestries. Study Design and Methods Marseilles Cord Blood Bank works with one university birth clinic caring for a culturally and sociologically diverse population. Stringent French legal restrictions apply to recording the geographic origin of parents. To circumvent this limitation and evaluate the contribution of newly banked cord blood units (CBUs) to increasing HLA diversity, we applied an algorithm that allows for the determination of parents' putative haplotypes and thus grossly deduce information on their ancestry. Generic resolution HLA-A, HLA-B, and allelic resolution HLA-DRB1 genotyping for 328 CBUs and 2691 unrelated donors (UDs) between January 2009 and May 2012 were performed. Enrichment from international CBU registry with nonreferenced generic HLA-A, HLA-B, and allelic HLA-DRB1 phenotypes was compared between CBUs identified with one or two non-European haplotypes and CBUs identified with two European haplotypes. Results Marseilles CBUs display an increased proportion of HLA antigens frequently expressed in African populations compared to UDs. Whereas 93% of 199 CBUs identified with one or two non-European haplotypes enrich international CBU registry, this result is reduced to 42% for the 129 CBUs identified with two European haplotypes. Conclusion This study supports a new method to assess HLA diversity. However, such an increased of HLA diversity raises questions about frequencies of CBUs released and clinical relevance from their uses.

International audience

BACKGROUND To assess the safety and potential efficacy of a standardized technique consisting of intra-articular injection of 10 cc of a homogeneous mixed product using autologous micro-fat and platelet rich plasma (PRP) (ratio 1:1) in the carpus or the fetlock joint of sport horses presenting degenerative joint disease (DJD). METHODS Eight sport horses with DJD confirmed by radiography and ultrasonography and causing lameness and the impossibility to compete were treated. PRP was prepared after a double centrifugation whereas micro-fat was harvested and purified using a closed system. The two products were connected and mixed by gentle back and forth shaking of the syringes to finally obtain 10 ml of an homogeneous mixed product. Follow up was performed from 5 to 10 months with assessment of AAEP lameness score and return to training and competition. RESULTS Nine joints were treated with significant improvement of the AAEP lameness score three months after the procedure (p = 0.021). Four horses returned to official competition between 5 to 10 months after the procedure (7.0±2.5) and three of them resumed intensive training between 5 to 9 months (6.3±2.3). No adverse event occurred. CONCLUSION This study is a first step in the development of innovative therapy for DJD which combines the potential chondrogenic differentiation of MSCs inside equine adipose tissue with the proliferative effect of growth factors present in PRP.

International audience

International audience; Objective. Impaired hand function greatly contributes to disability and reduced quality of life in SSc patients. Autologous adipose-derived stromal vascular fraction (ADSVF) is recognized as an easily accessible source of regenerative cells. We reported positive 6-month safety and efficacy results from an open-label clinical trial assessing s.c. injection of autologous ADSVF into the fingers in SSc patients. The objective of this report is to describe the effects at 12 months. Methods. Twelve females, mean age 54.5 years (S.D. 10.3), were assessed 1 year after ADSVF injection. Patients were eligible if they had a Cochin Hand Function Scale score >20/90. ADSVF was obtained from lipoaspirate using an automated processing system and subsequently injected into the s.c. tissue of each finger in contact with neurovascular pedicles in a one-time procedure. Endpoints were changes in hand disability and skin fibrosis, vascular manifestations, pain and quality of life at the 12 month follow-up. During the visit, patients estimated the benefit of the procedure with a specific self-completed questionnaire. Results. A significant decrease from baseline of 51.3% (P< 0.001) for Cochin Hand Function Scale score, 63.2% (P< 0.001) for RP severity and 46.8% (P = 0.001) for quality of life (Scleroderma Health Assessment Questionnaire) was observed. A significant improvement of finger oedema, skin sclerosis, motion and strength of the hands and of the vascular suppression score was also noted. The reduction in hand pain approached statistical significance (P = 0.052). The questionnaire revealed a benefit in daily activities, housework and social activities. Conclusion. ADSVF injection is a promising therapy and appears to have benefits that extend for at least 1 year.

The platelet-rich plasma (PRP) is an autologous biotherapy based on platelet-healing properties. Here, we developed a simple and reproducible PRP purification protocol based on two successive centrifugations. We evaluated different centrifugation speeds and time-storage durations on the platelet quantity and quality. Sterility and stability of our PRP homemade product were also performed. We prepared PRP from 54 healthy volunteers. We tested activation state, reactivity, and stability of platelets by flow cytometry using basal and adenosine diphosphate (ADP)-induced P-selectin expression markers; growth factor release after platelet activation by an enzyme-linked immunosorbent assay (ELISA); platelet aggregation capacity by aggregrometry assays; clot formation and retraction by thromboelastography; and platelet morphology by ultrastructural analysis. About 130 and 250 g successive speed centrifugations further concentrated platelets while preserving their bioactivity during 6 h (after that, platelet functions were significantly altered). In these conditions, we obtained a highly concentrated pure PRP product (with a low leukocyte count) suitable to study platelet properties. To avoid the loss of efficacy, we recommend injecting PRP under 3 h after preparation.

Grafted fat has many qualities of ideal filler; it is autologous, easily available, and naturally integrated into the host tissues. From a lipoaspirate, the adipose-tissue-derived stromal vascular fraction can be isolated, which is an excellent source of stem/stromal cells, endothelial progenitors, and immune cells. Fat grafting is being increasingly applied in autoimmune diseases, and this article focuses on systemic sclerosis, a rare autoimmune disease characterized by skin fibrosis and microvascular damage. The authors' approach of using fat graft in the face and adipose-tissue-derived stromal vascular fraction for hands is presented as innovative and promising therapy for patients with systemic sclerosis.

Several Cord Blood (CB) Bank studies suggested that ethnicity impaired CB unit (CBU) qualification. The Bone Marrow Donors Worldwide registries present an over-representation of unrelated donors (UD) from Northwestern European descent. This raises the question of equality of access to hematopoietic stem cells transplant, especially in the Mediterranean zone, which has taken in many waves of immigration. The aim of our study is to address whether, in the Marseille CB Bank, CBU qualification rate is impaired by geographic origin. The study compared biological characteristics of 106 CBU disqualified for total nucleated cell (TNC) count (dCBU) and 136 qualified CBU in relation to registry enrichment and haplotype origin. A high proportion (>80%) of both dCBU and CBU had at least one non-European haplotype and enrich CB and UD registries to a higher extent than those with two European haplotypes (P

Introduction Scleroderma is characterized by cutaneous manifestations that mainly affect the hands, arms and face. As of today, there is no treatment for fibrotic skin lesions of scleroderma. Previously we generated and validated a model of scleroderma-like skin sclerosis in nude mice, appropriate to inject human derived products. We showed that the subcutaneous injection of micro-fat (MF), purified and injected using small caliber cannulas, have anti-fibrotic and pro-angiogenic effects and appears more suitable for the treatment of skin lesions of scleroderma compared to the gold standard (Coleman’s technique or macro-fat). Here we compared the long-term efficacy of micro-fat “enriched” with other therapeutic products including the stromal vascular fraction (SVF) of fat and platelet-rich plasma (PRP) from blood in our murine model of scleroderma. Methods We used 72 nude mice in this study. We formed six experimental groups: Macro-fat, MF, SVF, PRP, MF + SVF, MF + PRP. This project has three phases: i) Induction of skin sclerosis by daily subcutaneous injections of bleomycin (BLM) for 4 weeks in nude mice; ii) Purification and injection of the different cell therapy products; iii) Histological analyses done 8 weeks post-injections. Results MF + SVF and MF + PRP significantly reversed dermal and epidermal sclerosis (P

International audience; BackgroundCord blood is an important source of stem cells. However, nearly 90% of public cord blood banks have declared that they are struggling to maintain their financial sustainability and avoid bankruptcy. The objective of this study is to evaluate how characteristics of cord blood units influence their utilization, then use this information to model the economic viability and therapeutic value of different banking strategies.MethodsRetrospective analysis of cord blood data registered between January 1st, 2009 and December 31st, 2011 in Bone Marrow Donor Worldwide. Data were collected from four public banks in France, Germany and the USA. Samples were eligible for inclusion in the analysis if data on cord blood and maternal HLA typing and biological characteristics after processing were available (total nucleated and CD34+ cell counts). 9,396 banked cord blood units were analyzed, of which 5,815 were Caucasian in origin. A multivariate logistic regression model assessed the influence of three parameters on the CBU utilization rate: ethnic background, total nucleated and CD34+ cell counts. From this model, we elaborated a Utilization Score reflecting the probability of transplantation for each cord blood unit. We stratified three Utilization Score thresholds representing four different banking strategies, from the least selective (scenario A) to the most selective (scenario D). We measured the cost-effectiveness ratio for each strategy by comparing performance in terms of number of transplanted cord blood units and level of financial deficit.ResultsWhen comparing inputs and outputs over three years, Scenario A represented the most extreme case as it delivered the highest therapeutic value for patients (284 CBUs transplanted) along with the highest financial deficit (USD 5.89 million). We found that scenario C resulted in 219 CBUs transplanted with a limited deficit (USD 0.98 million) that charities and public health could realistically finance over the long term. We also found that using a pre-freezing level of 18 x 108 TNC would be the most cost-effective strategy for a public bank.ConclusionOur study shows that a swift transition from strategy A to C can play a vital role in preventing public cord blood banks worldwide from collapsing.

Introduction: Patients with sickle cell disease (SCD) may develop a persistent mixed chimerism (MC) after hematopoietic stem cell transplantation (HSCT) associated with clinical control of the disease. In order to further investigate this condition we analyzed the chimerism in sorted myeloid and lymphoid subpopulations, and erythroid progenitors.Methods: Between 1990 and 2013, 112 sickle cell disease (SCD) patients underwent allogeneic HSCT after myeloablative conditioning. Among the 107 patients with available chimerism at 2 years, 55.1% had a MC, i.e. 9590nana6AA9111.893.83.13.10191223311337.5577AS15513.556.5412.60583471na4975604AS13210.65433.3nana5344444658323717AS1311.547.841.23.21.48575na9273837618A/Dpubjab8012.647.90nana887795959494.59912A/b0Thal579.69503291689698999910011AA5413.49703088781001009998985AA3410.978.38nana444641na5058432AS468.42468.1nana21709.45.43112.5na8AS1339.734.7632.401631121013089AS1539.552.145.920183026161117153AA3310.189.553.22.330631718231317

Background Combined immunodeficiencies (CIDs) form a heterogeneous group of inherited conditions that affect the development, function, or both of T cells. The treatment of CIDs with allogeneic hematopoietic stem cell transplantation (HSCT) is complicated by a high incidence of life-threatening infections and an increased risk of graft-versus-host disease (GVHD). Objective In view of the growing evidence that alloreactivity is mainly derived from human naive T cells, the selective depletion of naive T cells from allografts might constitute a way of reducing alloreactivity while maintaining memory T-cell responsiveness to pathogens. Methods Five consecutive patients with CIDs and chronic viral infections underwent an allogeneic, HLA-mismatched HSCT. Given the patients' infection status and the potential risk of severe GVHD in the mismatched setting, the CD34 − fraction of the allograft was depleted of naive T cells by using magnetic CD45RA beads. Results Engraftment occurred in 4 of the 5 patients. No severe GVHD occurred. In the 4 engrafted patients viral infections were cleared within 2 months of the HSCT, and both cellular and humoral immunity were re-established within a year of the HSCT. An early T-cell response against viral pathogens was documented in 2 patients. Conclusion The present pilot study shows that clinical-grade depletion of naive T cells from an allograft through the use of magnetic CD45RA beads seems to be a feasible and efficacious option for the treatment of patients with CIDs at high risk of GVHD, infection, or both in an HLA-mismatched setting.

The platelet-rich plasma (PRP) is an autologous biotherapy commonly used for its healing properties. Once activated, platelets released a real “cocktail” of growth factor and cytokines implied in numerous regenerative processes. However the impact of medical practices associated to PRP therapeutic use on platelets functionality remains poorly known. Objectives: we evaluated the in vitro effects of two commonly used local anesthetics (Xylocaine* and Naropin*) on PRP functionality. We also investigated the quantity and quality of PRP that passed through the smallest gauge needle commercialized. Materials and methods: PRP from 9 healthy volunteers were prepared using our previously described home made purification protocol. Platelet aggregation capacity was evaluated by aggregometry assays and the growth factor release was determined by ELISA after platelet activation. We also evaluated the platelet activation status, reactivity and stability of platelets by flow cytometry using the P-selectin expression marker. Results: the association of local anaesthetics with PRP injections resulted in a significant decrease of platelets functionality, assessed by their capacity of aggregating. Local anaesthetics did not interfere with the growth factor release. The different needle sizes and calibres tested for PRP injections did not influence the platelet functionality. Conclusions: the use of local anaesthetics to prevent pain during PRP injections could compromise the therapeutic potential of PRP. These results suggest using carefully local anaesthetics or limiting their use as often is possible. To minimize injection pain, we recommend using 30 G needles. These data will lead to clinical recommendations for painless and controlled PRP injections

International audience; BACKGROUND: New strategies are emerging in cord blood banking where focusing on birth clinics caring for a high number of mothers belonging to ethnic minorities could offer new possibilities for allotransplantation both for patients of European origin and for patients from ethnic minorities or mixed ancestries. STUDY DESIGN AND METHODS: Marseilles Cord Blood Bank works with one university birth clinic caring for a culturally and sociologically diverse population. Stringent French legal restrictions apply to recording the geographic origin of parents. To circumvent this limitation and evaluate the contribution of newly banked cord blood units (CBUs) to increasing HLA diversity, we applied an algorithm that allows for the determination of parents' putative haplotypes and thus grossly deduce information on their ancestry. Generic resolution HLA-A, HLA-B, and allelic resolution HLA-DRB1 genotyping for 328 CBUs and 2691 unrelated donors (UDs) between January 2009 and May 2012 were performed. Enrichment from international CBU registry with nonreferenced generic HLA-A, HLA-B, and allelic HLA-DRB1 phenotypes was compared between CBUs identified with one or two non-European haplotypes and CBUs identified with two European haplotypes. RESULTS: Marseilles CBUs display an increased proportion of HLA antigens frequently expressed in African populations compared to UDs. Whereas 93% of 199 CBUs identified with one or two non-European haplotypes enrich international CBU registry, this result is reduced to 42% for the 129 CBUs identified with two European haplotypes. CONCLUSION: This study supports a new method to assess HLA diversity. However, such an increased of HLA diversity raises questions about frequencies of CBUs released and clinical relevance from their uses.

Background: Autologous adipose tissue injection is used in plastic surgery for correction of localized tissue atrophy and has also been successfully offered for treatment of localized scleroderma. We aimed to evaluate whether patients with systemic sclerosis (SSc) and facial handicap could also benefit from this therapy. Methods: We included 14 patients (mean age of 53.8 ± 9.6 years) suffering from SSc with facial handicap defined by Mouth Handicap in Systemic Sclerosis Scale (MHISS) score more than or equal to 20, a Rodnan skin score on the face more than or equal to 1, and maximal mouth opening of less than 55 mm. Autologous adipose tissue injection was performed under local anesthesia using the technique of subcutaneous microinjection. The main objective of this study was an improvement of the MHISS score 6 months after the surgical treatment. Results: The procedure was well tolerated. We observed a mean decrease in the MHISS score of 10.7 points (±5.1; P < 0.0001) at 6 months (35% improvement). Secondary efficacy parameters assessing perioral skin sclerosis, maximum mouth opening, sicca syndrome, and facial pain significantly improved at 3 and 6 months postsurgery. At a 6-month follow-up, 75% of patients were satisfied or very satisfied of the adipose tissue microinjection therapy. Conclusions: Our study suggests that subcutaneous perioral microfat injection in patients with SSc is beneficial in the treatment of facial handicap, skin sclerosis, mouth opening limitation, sicca syndrome, and facial pain. Thus, this minimally invasive approach offers a new hope for face therapy for patients with SSc.

Introduction Les arthroses du poignet sont invalidantes, lourdes de consequences socio-economiques et beneficient a l’heure actuelle de traitements medicaux d’efficacite limitee et de traitements chirurgicaux lourds. En medecine regenerative, les injections intra-articulaires de plasma riche en plaquettes n’ont pas ete rapportees dans le poignet alors qu’elles connaissent un essor considerable dans le genou. Les injections de tissu adipeux intra-articulaires n’ont pas non plus ete rapportees. En medecine veterinaire, les chevaux de course sont soumis a des pathologies articulaires degeneratives des membres anterieurs, en raison d’une usure prematuree et de traumatismes repetes. Le traitement de l’osteoarthropathie degenerative du cheval est devenu une priorite de recherche. Le cheval, qui possede un membre anterieur anatomiquement comparable au membre superieur de l’homme, est un excellent modele preclinique pour le traitement des pathologies articulaires du carpe et de l’articulation metacarpophalangienne. Nous presentons une serie du 8 cas de chevaux de competition atteints d’osteoarthropathie degenerative, presentant cliniquement une boiterie ayant entraine l’arret de la competition, traites par un melange de micrograisse autologue et de plasma riche en plaquettes injecte en intra-articulaire dans le carpe ou la metacarpophalangienne, avec un recul de 3 mois. Materiel et methodes Les criteres d’inclusion etaient la presence d’une arthropathie severe confirmee par des modifications radiographiques et echographiques, et une boiterie clinique superieure ou egale a 1/5. Le protocole operatoire etait realise en quatre temps sur un cheval sedate : prelevement sanguin et preparation du PRP, prelevement de tissu adipeux et purification, preparation du melange et reinjection intra-articulaire. Resultats Aucun effet indesirable n’a ete rapporte. Apres 3 mois, 7 chevaux sur 8 montraient un meilleur confort de marche. Sept d’entre eux ont repris leur activite anterieure, dont 4 en competition et 3 en entrainement. Un a ete reforme au bout de 3 mois. Aucun cheval n’est parti a l’abattoir. Discussion Malgre un faible nombre de cas, cette etude montre de resultats encourageants sur la tolerance et l’efficacite des injections intra-articulaires de produits de therapie cellulaire chez le cheval de competition. Devant les similitudes anatomiques au membre anterieur du cheval et de l’homme et la physiopathologie similaire des lesions articulaires degeneratives, ces resultats nous encouragent a envisager des applications cliniques chez l’homme.

International audience; IMPORTANCE: Wiskott-Aldrich syndrome is a rare primary immunodeficiency associated with severe microthrombocytopenia. Partially HLA antigen-matched allogeneic hematopoietic stem cell (HSC) transplantation is often curative but is associated with significant comorbidity. OBJECTIVE: To assess the outcomes and safety of autologous HSC gene therapy in Wiskott-Aldrich syndrome. DESIGN, SETTING, AND PARTICIPANTS: Gene-corrected autologous HSCs were infused in 7 consecutive patients with severe Wiskott-Aldrich syndrome lacking HLA antigen-matched related or unrelated HSC donors (age range, 0.8-15.5 years; mean, 7 years) following myeloablative conditioning. Patients were enrolled in France and England and treated between December 2010 and January 2014. Follow-up of patients in this intermediate analysis ranged from 9 to 42 months. INTERVENTION: A single infusion of gene-modified CD34+ cells with an advanced lentiviral vector. MAIN OUTCOMES AND MEASURES: Primary outcomes were improvement at 24 months in eczema, frequency and severity of infections, bleeding tendency, and autoimmunity and reduction in disease-related days of hospitalization. Secondary outcomes were improvement in immunological and hematological characteristics and evidence of safety through vector integration analysis. RESULTS: Six of the 7 patients were alive at the time of last follow-up (mean and median follow-up, 28 months and 27 months, respectively) and showed sustained clinical benefit. One patient died 7 months after treatment of preexisting drug-resistant herpes virus infection. Eczema and susceptibility to infections resolved in all 6 patients. Autoimmunity improved in 5 of 5 patients. No severe bleeding episodes were recorded after treatment, and at last follow-up, all 6 surviving patients were free of blood product support and thrombopoietic agonists. Hospitalization days were reduced from a median of 25 days during the 2 years before treatment to a median of 0 days during the 2 years after treatment. All 6 surviving patients exhibited high-level, stable engraftment of functionally corrected lymphoid cells. The degree of myeloid cell engraftment and of platelet reconstitution correlated with the dose of gene-corrected cells administered. No evidence of vector-related toxicity was observed clinically or by molecular analysis. CONCLUSIONS AND RELEVANCE: This study demonstrated the feasibility of the use of gene therapy in patients with Wiskott-Aldrich syndrome. Controlled trials with larger numbers of patients are necessary to assess long-term outcomes and safety.

International audience; Introduction: Hand involvement confers a substantial handicap in work and daily activities in patients with Systemic sclerosis (SSc). Autologous adipose-derived stromal vascular fraction is as an easily accessible source of cells with regenerative effects. We previously performed a phase I open-label clinical trial (NTC01813279) assessing the safety of subcutaneous injection of autologous adipose-derived stromal vascular fraction. Six and 12-month data have been reported. As patients were followed in our medical centre, we report their longer-term outcome beyond the end of the trial.Patients and method: Twelve females, mean age 54.5 ± 10.3 years, initially enrolled in the clinical trial were assessed during a scheduled medical care, which took place between 22 and 30 months after treatment.Results: Multiple patient-reported outcomes showed sustained improvement, in comparison with the assessment performed just before surgery: 62.5% in the Cochin Hand Function Scale, 51.1% in the Scleroderma Health Assessment Questionnaire, 33.1% in hand pain, and 88.3% in the Raynaud Condition Score. A decrease in the number of digital ulcers number was noted. Mobility, strength and fibrosis of the hand also showed improvement. None of the 8 patients who had previously received iloprost infusion required new infusion.Conclusion: Despite the limits of an open label study, the data are in favour of the long-term safety of the adipose-derived stromal vascular fraction injection. Two randomized double blind, placebo-controlled trials of this therapeutic agent are ongoing in the USA (NCT02396238) and in France (NCT02558543) and will help determine the place of this innovative therapy for SSc patients.